An understanding of the mechanisms and pathways that mediate cardiac myocyte death is critical for the development of pharmacological therapies to limit heart damage during myocardial infarction (“heart attack”). We have completed the first unbiased cell-based screen of the MLSMR library consisting of ~360,000 compounds to identify small molecule probes that protect cardiac myocytes against oxidative and metabolic stresses, both precipitants of cell death during myocardial infarction. Oxidative stress was modeled with hydrogen peroxide treatment, and metabolic stress by inhibition of glycolysis using 2-deoxyglucose (2-DG). Since cardiac myocytes in vivo are terminally differentiated cells that cannot be expanded to the numbers needed for these studies, two cardiac myocyte model cell lines were used: H9c2 cells, derived fetal rat heart; and HL-1 cells, derived from the hearts of mice with cardiac-specific expression of an SV40 T-antigen transgene. Based on the activities of compounds in each of these cell types in response to each of these death stimuli (4 assays), 10 scaffolds were prioritized for reorder of their powders and available analogs for “analog-by-catalog” (ABC) testing. Two scaffolds advanced through multiple rounds of analog synthesis and structure-activity studies to result in the first probe molecule ML330. ML330 protects both H9c2 and HL-1 cell lines (EC50 0.4–4.0 μM & Emax 42–99 %) from both oxidative and metabolic stress induced by hydrogen peroxide and 2-DG, respectively. A second probe ML331, was identified preferentially protects both cell lines (EC50 0.6–9.0 μM & Emax 39–74 %) from metabolic stress induced by 2-DG. Probe molecules ML330 and ML331 will be useful to map and understand the critical cell death pathways in cardiac myocytes and lead to the development of a pharmacological agent to limit heart damage during myocardial infarction in humans. The probe molecules may also find broad utility in studies to understand relationships that regulate connections between various cell death programs, such as apoptosis and necrosis. Moreover, these findings may also extend to other ischemic syndromes such as stroke.